C-2015: Experimental Study of deglycerolization of red blood cells with a dilution–filtration system

Abstract

Freezing of red blood cells (RBCs) is an important application of cryopreservation. As a primary cryoprotectant of RBCs, glycerol must be removed from the thawed previously frozen RBCs prior to transfusion. The most commonly traditional method for deglycerolization is centrifugation, however, this has limitations in time consumption, operational complexity and high cost. Recently, a novel method with a better ability to remove large molecular substances called dilution-filtration was reported in the literature. The method involves pure ultrafiltration for separation, which essentially differentiates it from centrifugation. In this work, an in-depth experiment was done for deglycerolization of red blood cells. In order to achieve this goal, a fully automatic instrument based on the dilution–filtration theory has been developed. Also, the experiment requests to measure some basic haematological parameters of deglycerolized red blood cells, such as red blood cells count, hematocrit, residual glycerol, supernatant hemoglobin and extracellular potassium levels. All these experimental measurement parameters had to meet the blood transfusion requirements that established by the American Association of Blood Banking (AABB). Moreover, a high efficiency and low cost separation membrane product was determined by utilizing dialyzer, blood filter and plasma filter independently during the deglycerolization. Finally, the whole experiment process was carried out in the local blood center. Meanwhile, to ensure our experiment results could be highly persuasive, operations personnel were asked to abide a recommended quality control which was adapted from a standard operating procedure made by the Naval Blood Research Laboratory (NBRL). Taken together, the dilution-filtration system was proved to be efficient and feasible for removing glycerol from the cryopreserved red blood cells. Furthermore, with the ultimate success of industrial operations, it is expected to change centrifugation trades in the deglycerolized field in the future.